Following are the properties of ideal fixatives (buffered formalin)
An ideal fixative prevents autolysis and bacterial decomposition in the tissue.
- It preserves the tissue in its natural state.
- Fix all tissue components at their position.
- Makes the cellular component insoluble to the reagent used in tissue processing.
- Preserve the tissue volume.
- Avoid excessive hardness of tissue.
- Enhance tissue staining(H and E stain) process.
- It should be non-toxic and non-allergic to the user.
- Good fixative should be cheap.
Classification of chemical fixatives in the Histopathology laboratory
Fixatives are classified into three groups.
- Tissue fixatives
- Histochemical fixatives
- Cytology fixatives
What are the tissue fixative examples?
These fixatives are specially used for tissue fixation. Following fixatives used for tissue
- Buffered Formal saline
- Zenker Solution
- Buffered Glutaraldehyde
What are Histochemical fixative names?
These fixatives are specially used for histochemical fixation. Following fixatives used
- Cold acetone
- Absolute Alcohol
- Formal Saline
Types of cytological fixatives and examples
These fixatives are used for cytological specimens. Following fixatives used in the cytology
- Ether
- Methanol
- Ethanol
10 most common types of fixative used in Histopathology
Two fixatives are commonly used
- 10% Routine Formalin
- 10% Buffered Formalin
- Ethyl Alcohol
- Mercuric Chloride
- Picric Acid
- Chromic Acid
- Potassium Dichromate
- Osmic Acid (Osmium Tetra Oxide)
- B-5 Fixative
- Zinker Solution
5 Properties of 10% routine formalin histopathology
- It is prepared with a 40% w/w solution of formaldehyde gas in water.
- Routine formalin is used at 10% or 15% V/V in normal saline or calcium chloride solution.
- It does not precipitate PROTEIN. But combine with NH2 to form an insoluble gel.
- Preserves all components including fats. Phospholipid insoluble in fat solvents.
- Cheapest and most popular fixative in histology.
What is the disadvantage of 10% routine formalin fixative?
Routine formalin has acidic PH. This results in haematin crystal formation during fixation. These crystals also affect tissue staining(Hematoxylin and eosin).
10 neutral buffered formalin composition
- Normal saline 100 ml
- Pure formalin 10 ml
- Sodium dihydrogen phosphate 0.4 g
- Disodium hydrogen phosphate 0.65 g
Note: 10% buffered formalin is prepared after mixing all these substances.
6 Advantages of 10% neutral buffered formalin fixative
Following are the uses or advantages of a 10 percent neutral buffer
- Tissue can be preserved in it for a longer time.
- There is no hardening of the tissue.
- Tissue sectioning is easy.
- No haematin crystal is formed.
- It enhances tissue staining.
- A number of stains can be used on such preserved tissue.
Fixative of choice and Target table (Do, don’t)
The following fixatives are used as a fixative of choice for protein, lipid, and enzyme.
Fixative of choice | Target | To be avoided |
Frozen section | Enzyme | Chemical fixative |
Buffered formalin | Protein | Osmium tetraoxide |
Alcohol fixative | Nucleic acid | Aldehyde Fixative |
Alcohol-based fixative | Glycogen | Osmium tetraoxide |
Frozen section | Lipid | Alcohol, formalin |
Frozen Section | Mucopolysaccharide | Chemical Fixative |